ArticleMicrobiologyOpen2020
Pentaplex real-time PCR for differential detection of Yersinia pestis and Y. pseudotuberculosis and application for testing fleas collected during plague epizootics.
Article in MicrobiologyOpen, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed, 17 citations in OpenAlex.
- Establishment and application of a quadruple target real-time PCR assay for detecting Yersinia pestis.PloS one · 2026Article
- Ebselen, a promising host-directed therapeutic option againstFrontiers in microbiology · 2026Article
- An ELISA-like sensitive and visual detection system targetingJournal of clinical microbiology · 2025Article
- Development and evaluation of a triplex real-time PCR assay for enhanced plague diagnostics in Madagascar.PLoS neglected tropical diseases · 2025Article
- Multiplex PCR System for the Diagnosis of Plague.Current molecular medicine · 2025Review
- 125 years of the plague in Brazil: lessons learnt, historical insights and contemporary challenges.Memorias do Instituto Oswaldo Cruz · 2025Article
- Development and evaluation of a multi-target droplet digital PCR assay for highly sensitive and specific detection of Yersinia pestis.PLoS neglected tropical diseases · 2024Article
- Applications of polymerase chain reaction-based methods for the diagnosis of plague (Review).Experimental and therapeutic medicine · 2022Review
- A Novel Loop-Mediated Isothermal Amplification Assay for Rapid Detection ofFrontiers in microbiology · 2022Article
- No evidence for enzootic plague within black-tailed prairie dog (Cynomys ludovicianus) populations.Integrative zoology · 2021Article
- Obtaining Specific Sequence Tags forPathogens (Basel, Switzerland) · 2021Article
- Highly Specific and Sensitive Detection ofFrontiers in microbiology · 2021Article
- Pentaplex real-time PCR for differential detection of Yersinia pestis and Y. pseudotuberculosis and application for testing fleas collected during plague epizootics.MicrobiologyOpen · 2020Article
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Authors and funding
8 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Upon acquiring two unique plasmids (pMT1 and pPCP1) and genome rearrangement during the evolution from Yersinia pseudotuberculosis, the plague causative agent Y. pestis is closely related to Y. pseudotuberculosis genetically but became highly virulent. We developed a pentaplex real-time PCR assay that not only detects both Yersinia species but also differentiates Y. pestis strains regarding their plasmid profiles. The five targets used were Y. pestis-specific ypo2088, caf1, and pst located on the chromosome, plasmids pMT1 and pPCP1, respectively; Y. pseudotuberculosis-specific chromosomal gene opgG; and 18S ribosomal RNA gene as an internal control for flea DNA. All targets showed 100% specificity and high sensitivity with limits of detection ranging from 1 fg to 100 fg, with Y. pestis-specific pst as the most sensitive target. Using the assay, Y. pestis strains were differentiated 100% by their known plasmid profiles. Testing Y. pestis and Y. pseudotuberculosis-spiked flea DNA showed there is no interference from flea DNA on the amplification of targeted genes. Finally, we applied the assay for testing 102 fleas collected from prairie dog burrows where prairie dog die-off was reported months before flea collection. All flea DNA was amplified by 18S rRNA; no Y. pseudotuberculosis was detected; one flea was positive for all Y. pestis-specific targets, confirming local Y. pestis transmission. Our results indicated the assay is sensitive and specific for the detection and differentiation of Y. pestis and Y. pseudotuberculosis. The assay can be used in field investigations for the rapid identification of the plague causative agent.
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